2004 Fiscal Year Final Research Report Summary
Development of serum diagnosis kit against osteosarcoma and other tumor.
| Project/Area Number |
14571400
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | Nara Medical University |
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Principal Investigator |
MAEDA Manabu Nara Medical University, Emergency Unit, Assistant Professor, 医学部, 助手 (80311926)
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| Co-Investigator(Kenkyū-buntansha) |
MII Nobuo Nara Medical University, Orthopedic surgery, Professor emeritus, 医学部, 名誉教授 (70145845)
HATANO Osamu Nara Medical University, Anatomy, Lecturer, 医学部, 講師 (40164850)
OKARU Haruhiko Dainippon Pharmacy Ltd., Department of Laboratory products, director, ラボラトリープロダクツ部・研究開発部, 部長
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| Project Period (FY) |
2002 – 2004
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| Keywords | serum diagnosis / monoclonal antibody / tumor / PCNA |
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| Research Abstract |
We made attention to construct diagnosis kit from sera this year. To make the kit, we laid emphasis to make monoclonal antibodies. And the more the most important thing to make monoclonal antibodies, is to prepare antigen. We tried to purify antigen from rabbit thymus acetone powder with simple method. For the purification of protein, various kinds of columns were tested and found out to use an ion exchange chromatography is the best way. We purchased rabbit thymus acetone powder and partial purified PCNA related protein. To check the protein if it is PCNA related protein or not, we used polyclonal antibody against PCNA with commercially available. The antibody recognized one strong band with the molecular weight between 35KD and 40 KD. Because PCNA protein shows 36〜37KD. But unfortunately, the antibody also recognized another protein with molecular weight of more than 100KD. Next step was to remove the additional band. We cut gels appropriate area and extract protein by using electro-phoresis. This area contains only one band which reacted with the antibody. We next prepared monoclonal antibodies using this protein by inject to abdominal cavity of several mice with adjuvant. After make sure that mouse express antibody against the purified protein, we removed spleen and hybridized with mouse myeloma cell line of X63. We did subcloning twice. Finally we got two good hybridoma cell lines. These cell lines express IgM type monoclonal antibodies. We now repeat making hybridomas with express IgG type antibodies.
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Research Products
(12 results)
